陕西师范大学学报（自然科学版）2017，Vol.45Issue(5)：72-77,6.DOI:10.15983/j.cnki.jsnu.2017.05.355
解脂亚罗酵母中HMGR基因的生物信息学分析及催化域部分表达
Bioinformatics analysis of HMGR gene and partial expression of catalytic domain in Yarrowia lipolytica
摘要
Abstract
Bioinformatics analysis of the HMGR of rate-limiting enzyme in the mevalonate pathway from Yarrowia lipolytica was carried out.The activity domain (tHMG,amino acids 441～875) was cloned and co-expressed with the labeled molecule GFP.Bioinformatics analysis showed that the HMGR gene can encode a hydrophobic labile protein containing 999 amino acids.The first 500 amino acid peptide chains contain more transmembrane region,the molecular weight was 254.93 kDa and theoretical pI was 4.77.The active domain contains 506 amino acids with a molecular weight of 127.74 kDa and a theoretical isoelectric point of 4.93,which contains the nontransmembrane region.The secondary structures of the two proteins (HMGR and tHMG) are mainly α-helix and random coil,and their tertiary structures are in the state of homologous tetramer.The yeast cells expressing tHMG-GFP were observed at the excitation wavelength of 488 nm,and the green fluorescence was found.After overexpression of catalytic domain (tHMG),the enzyme activity was 1.14 U/mg,which was 62.7％ higher than that of the control.The results showed that the catalytic domain of HMGR could be expressed independently and it can increase the enzyme activity of HMGR in the cells.关键词
解脂亚罗酵母/甲羟戊酸途径/3-羟基-3-甲基戊二酰辅酶A还原酶/生物信息学分析/融合表达Key words
Yarrowia lipolytica/mevalonate pathway/3-hydroxy-3-methylglutaryl-CoA reductase/bioinformatics analysis/fusion expression分类
农业科技引用本文复制引用
屈玉玲,谭思远,董桂茹,苏安平,孟永宏..解脂亚罗酵母中HMGR基因的生物信息学分析及催化域部分表达[J].陕西师范大学学报（自然科学版）,2017,45(5):72-77,6.基金项目
陕西省科技统筹项目(2015KTCQ02-07) （2015KTCQ02-07）
国家重点科技研发计划(2017YFD0400700) （2017YFD0400700）
